Quantification of the histochemical staining for carbonyles and DNA using 3-hydroxy-2-naphthoic acid hydrazide and fast blue B.
Fixed cells and tissues pretreated with 4-hydroxynonenal were used as models for the histochemical demonstration of protein bound aldehydic groups. The aldehydes were stained with both a modification of the 2,4-dinitrophenylhydrazine method (2,4-DNPH) and the optimized staining using 3-hydroxy-2-naphthoic acid hydrazide and Fast blue B (NAH-FB). A correlation has been found between the specific microphotometric mean integrated maximum absorbance values of cells and tissues stained with 2,4-DNPH and with NAH-FB (cc = 0.999). The maximum absorbance measured after 2,4-DNPH-staining (epsilon 367 = 21,000) were 1.893 +/- 0.072 (P less than 0.01) times that of NAH-FB-staining at 550 nm. Microphotometrically determined DNA-values of different cells stained with the NAH-FB-DNA-method correlated with those determined with methods of analytical biochemistry and published by other authors.
Nöhammer
G
G
Institute of Biochemistry, Karl-Franzens-University Graz, Austria.
D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism / Q000473:pathology; D004247:DNA / Q000378:metabolism*; D003979:Diazonium Compounds / Q000378:metabolism*; D006651:Histocytochemistry; D008099:Liver / Q000166:cytology; D008832:Microchemistry / Q000379:methods; D009284:Naphthols / Q000378:metabolism*; D010659:Phenylhydrazines / Q000378:metabolism; D010783:Photometry / Q000379:methods; D011506:Proteins / Q000378:metabolism; D051381:Rats
D000447:Aldehydes; D003979:Diazonium Compounds; D009284:Naphthols; D010659:Phenylhydrazines; D011506:Proteins; C004787:2,4-dinitrophenylhydrazine; C020747:Fast Blue B; C010589:3-hydroxy naphthoic acid hydrazide; D004247:DNA; C027576:4-hydroxy-2-nonenal